The essential and non-essential genes of Bovine herpesvirus 1.
Bovine herpesvirus 1 (BoHV-1) is an economically important pathogen of cattle associated with respiratory and reproductive disease. To further develop BoHV-1 as a vaccine vector, a study was conducted to identify the essential and non-essential genes required for in vitro viability. Random-insertion mutagenesis utilizing a Tn5 transposition system and targeted gene deletion were employed to construct gene disruption and gene deletion libraries, respectively, of an infectious clone of BoHV-1. Transposon insertion position and confirmation of gene deletion were determined by direct sequencing. The essential or non-essential requirement of either transposed or deleted open reading frames (ORFs) was assessed by transfection of respective BoHV-1 DNA into host cells. Of the 73 recognized ORFs encoded by the BoHV-1 genome, 33 were determined to be essential and 36 to be non-essential for virus viability in cell culture; determining the requirement of the two dual copy ORFs was inconclusive. The majority of ORFs were shown to conform to the in vitro requirements of BoHV-1 homologues encoded by human herpesvirus 1 (HHV-1). However, ORFs encoding glycoprotein K (UL53), regulatory, membrane, tegument and capsid proteins (UL54, UL49.5, UL49, UL35, UL20, UL16 and UL7) were shown to differ in requirement when compared to HHV-1-encoded homologues.
Robinson
Karl E
KE
School of Veterinary Science, University of Queensland, St Lucia, Brisbane, QLD, Australia.|Department of Primary Industries and Fisheries, St Lucia, Brisbane, QLD, Australia.
Meers
Joanne
J
School of Veterinary Science, University of Queensland, St Lucia, Brisbane, QLD, Australia.
Gravel
Jennifer L
JL
Department of Primary Industries and Fisheries, St Lucia, Brisbane, QLD, Australia.
McCarthy
Fiona M
FM
Department of Primary Industries and Fisheries, St Lucia, Brisbane, QLD, Australia.
Mahony
Timothy J
TJ
Department of Primary Industries and Fisheries, St Lucia, Brisbane, QLD, Australia.
D000818:Animals; D002417:Cattle; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015723:Gene Library; D020043:Genes, Essential; D005814:Genes, Viral; D016679:Genome, Viral; D007242:Herpesvirus 1, Bovine / Q000235:genetics* / Q000276:immunology; D018259:Herpesvirus 1, Human / Q000235:genetics; D006801:Humans; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D014162:Transfection; D014765:Viral Vaccines; D014779:Virus Replication / Q000235:genetics
D004279:DNA, Viral; D014765:Viral Vaccines